PROTEINS IN LEAD-INDUCED NUCLEAR INCLUSION BODIES

铅诱发的核包涵体中的蛋白质

• 批准号: 3249745
• 负责人: KEITH R SHELTON
• 金额: $ 12.2万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-08-01 至 1987-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3249745
• 关键词: autoradiography; electrofocusing; electron microscopy; environmental toxicology; gel electrophoresis; gene expression; genes; genetic regulation; inclusion body; lead poisoning; molecular cloning; monoclonal antibody; nucleic acid sequence; phase contrast microscopy; radionuclides; renal cortex; tissue /cell culture

Lead poisoning induces nuclear bodies, composed of lead and protein, in many animals including man. These bodies are prominent in renal tubular lining cells. There are well known clinical effects of lead poisoning, but neither the function nor the effect of the nuclear inclusion bodies is known. They may reflect a regulated process which decreases the level of soluble lead. Their formation is blocked in cell culture by inhibitors of RNA and protein synthesis, suggesting that inducible proteins are involved. The inclusion bodies contain a prominent protein which has not been detected elsewhere, suggesting that it is either induced or sequestered by lead. In order to understand inclusion body formation and function, it is necessary to relate the metabolism of these proteins to lead exposure and to study the interactions of the proteins with lead. The chemical properties of the inclusion bodies impose severe restraints on their study. A cell culture system will be used in correlating synthesis of inclusion body proteins with lead exposure. Further, the synthesis of the cognate mRNA's will be determined by an in vitro translation assay in order to confirm induction by lead. The tendency of these proteins to bind lead will be studied by exposure of purified, bound protein to soluble radioactive lead. Monoclonal antibodies will be prepared and used for assaying and purifying the proteins and in characterizing their intracellular location(s). Complementary DNA clones will be isolated in order that regulation of gene activity can be studied quantitatively in various cells and under various stimuli such as lead, other metals, and other traumatic effects such as heat. The effects of lead on cells have been difficult to study at the molecular level. However, the application of recently developed procedures to a study of the long recognized nuclear inclusion bodies provides a unique opportunity to study this problem.

铅中毒会诱发由铅和蛋白质组成的核体 许多动物，包括人。 这些小体在肾小管中突出 衬里细胞。 铅中毒的临床效应众所周知，但是 核包涵体的功能和作用都不是 已知。 它们可能反映了一个降低水平的受监管的过程 可溶性铅。 它们的形成在细胞培养物中被以下抑制剂阻断： RNA 和蛋白质合成，表明诱导蛋白是 涉及。 包涵体含有一种突出的蛋白质，该蛋白质没有 在其他地方检测到，表明它是诱导的或 被铅隔离。 为了了解包涵体的形成和 功能，有必要将这些蛋白质的代谢与 铅暴露并研究蛋白质与铅的相互作用。 这 包涵体的化学性质对 他们的学习。 细胞培养系统将用于关联合成 铅暴露的包涵体蛋白。 进一步地，合成 同源 mRNA 将通过体外翻译测定来确定 为了确认铅的诱导。 这些蛋白质结合的倾向 将通过将纯化的结合蛋白质暴露于可溶性蛋白质来研究铅 放射性铅。 单克隆抗体将被制备并用于 测定和纯化蛋白质并表征它们 细胞内位置。 互补 DNA 克隆将被分离到 为了能够定量研究基因活性的调节 各种细胞和各种刺激下，如铅、其他金属和 其他创伤性影响，例如热。 铅对细胞的影响有 很难在分子水平上进行研究。 然而，应用程序 最近开发的程序来研究长期公认的核 包容性机构为研究这个问题提供了独特的机会。